Effect of NDC80 in human hepatocellular carcinoma

AIM To investigate the role of nuclear division cycle(NDC)80in human hepatocellular carcinogenesis.METHODS NDC80 gene expression was analyzed by real-time reverse transcription polymerase chain reaction in 47paired hepatocellular carcinoma(HCC)and adjacent tissues.The HCC cell line SMMC-7721 was transfected with lentivirus to silence endogenous NDC80 gene expression,which was confirmed by real-time polymerase chain reaction and western blotting.The effects of NDC80silencing on SMMC-7721 cell proliferation were evaluated by Cellomics Array Scan VTI imaging.Cell cycle analysis and apoptosis were detected with flow cytometry.Colony formation was assessed by fluorescence microscopy.RESULTS NDC80 expression levels in HCC tissues were significantly higher than those in the adjacent tissues.Functional studies demonstrated that NDC80 silencing significantly reduced SMMC-7721 cell proliferation and colony formation.Knockdown of NDC80 resulted in increased apoptosis and cell cycle arrest at S-phase.NDC80 contributed to HCC progression by reducing apoptosis and overcoming cell cycle arrest. CONCLUSION Elevated expression of NDC80 may play a role in promoting the development of HCC.

ANGPTL2 expression in gastric cancer tissues and cells and its biological behavior

AIM To explore expression of angiopoietin-like protein 2(ANGpT L2) and its effect on biological behavior such as proliferation and invasiveness in gastric cancer. METHODS Western blotting was used to detect expression of ANGp TL2 in 60 human normal gastric tissues, 60 human gastric cancer tissues and gastric cell lines including GES-1, N87, SGC7901, BGC823 and pA MC82. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) and Transwell assay were used to detect the proliferation and invasive ability of gastric cancer cells. RESULTS Compared to normal tissues, ANGp TL2 protein levels were significantly upregulated in gastric tissues, and this level was closely correlated with gastric tumor grade, clinical stage and lymph node metastasis. Compared to GES-1 cells, ANGpT L2 mR NA and protein levels were significantly increased in gastric cancer cells including N87, SGC7901, BGC823 and p AMC82. The expression of ANGpT L2 in highly malignant gastric cancer cell lines BGC823 and pA MC82 was significantly higher than in low malignancy gastric cancer cell lines N87 and SGC7901. MTT and Transwell experiments indicated that the proliferation rate and invasive ability of stable overexpressed gastric cancer cells was faster than in cells transfected with Lv-NC and blank controlcells, and the invasive ability of stable overexpressed gastric cancer cells was higher than that of cells transfected with Lv-NC and blank control cells.CONCLUSION ANGp TL2 contributed to proliferation and invasion of gastric cancer cells. In clinical treatment, ANGpT L2 may become a new target for treatment of gastric cancer.

Lycopene modulates cellular proliferation, glycolysis and hepatic ultrastructure during hepatocellular carcinoma

AIM To investigate the effect of lycopene extracted from tomatoes(LycT) on ultrastructure, glycolytic enzymes, cell proliferation markers and hypoxia during N-Nitrosodiethylamine(NDEA)-induced hepatocarcinogenesis. METHODS Female BALB/c mice were randomly divided into four groups: The Control, NDEA(200 mg NDEA/kg b.w. given i.p.), LycT(5 mg/kg b.w. given orally on alternate days) and Lyc T + NDEA group. The m RNA and protein expression of various cell proliferation markers(PCNA, Cyclin D1, and p21) were assessed by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively. The ultrastructure of hepatic tissue was analyzed using scanning and transmission electron microscopy. The enzymatic activity of glycolytic enzymes was estimated using standardized protocols, while glucose-6-phosphate dehydrogenase activity level was estimated using a kit obtained from Reckon Diagnostic P. Ltd.(India). RESULTS Uncontrolled proliferation in the liver of NDEA(P ≤ 0.001) mice was evident from the high expression of cell-proliferation associated genes(PCNA, Cyclin D1, and p21) when compared to control and Lyc T mice. In addition, enhanced activities of hexokinase, phosphoglucoisomerase, aldolase, glucose-6-phosphatedehydrogenase and hypoxia-inducible factor-1α were observed in NDEA mice as compared to control(P ≤ 0.001) and Lyc T(P ≤ 0.001) mice. The alterations in hepatic ultrastructure observed in the NDEA group correlated with the changes in the above parameters. Lyc T pre-treatment in NDEA-challenged mice ameliorated the investigated pathways disrupted by NDEA treatment. Moreover, hepatic electron micrographs from the Lyc T + NDEA group showed increased macrophages, apoptotic bodies and well-differentiated hepatocellular carcinoma(HCC) in comparison to undifferentiated HCC as observed in the NDEA treated group. CONCLUSION This study demonstrates that dietary supplementation with Lyc T has a multidimensional role in preventing HCC development.